BRPI0612309A2 - antiviral compounds and methods - Google Patents

Abstract

ANTI VIAL COMPOUNDS AND METHODS. The present invention relates to novel compounds and compositions having antiviral activity. The invention also relates to methods for the therapeutic or prophylactic treatment of viral infections in mammals.

Description

Report of the Invention Patent for "ANTIVIAL COMPOUNDS AND METHODS".

FIELD OF INVENTION

The present invention relates to novel compounds and compositions having antiviral activity. The invention also relates to methods for delaying, reducing or otherwise inhibiting viral growth and / or functional activity.

BACKGROUND OF THE INVENTION

Currently, there is a great need for the development of new treatments that are effective against viral infections, particularly against viral infections that are associated with high morbidity and mortality, and impact on populations of considerable size. Currently available treatments are inadequate or ineffective in large proportions of infected patients.

A large number of viruses contribute to the gathering of significant human pathogens. Examples of these include the Lentivirus and Flavivirus family viruses, for example, HIV, Hapatite C virus (HCV), Dengue virus, and the like.

To refine the prospect of treatment and prevention of viral infections, and to deal with the advancement of viral evolution, there is a necessary advance to identify molecules capable of inhibiting various aspects of viral life cycle. A number of such compounds are disclosed in PCT / AU2004 / 000866. However, there is still a need for new compounds and additional agents with antiviral activity.

It is an object of the present invention to overcome or improve at least one of the disadvantages of the prior art, or to provide a useful alternative.

Any discussion of the prior art throughout any descriptive report should not be considered as an admission that such prior art is widely known or part of general knowledge at the time.

SUMMARY OF THE INVENTION The inventors have surprisingly found that certain compounds falling under the classification of substituted acylguanidines have antiviral activity against viruses from a range of different virus families. A number of such compounds are disclosed in PCT / AU2004 / 000866, incorporated in their entirety herein by reference.

The present invention relates to certain novel antiviral compounds that fall within the classification of substituted acylguanidines.

According to a first aspect, the present invention provides compound of Formula I:

<formula> formula see original document page 3 </formula>

in which

R1 is phenyl, substituted phenyl, naphthyl, substituted naphthyl, or R1 is selected from

<formula> formula see original document page 3 </formula>

It is independently halogen, alkyl, halo or polyhaloalkyl;

Q is independently hydrogen, especially methoxy alkoxy, especially methyl alkyl, cycloalkyl, thienyl, furyl, pyrazoyl, substituted pyrazoyl, pyridyl, substituted pyridyl, phenyl, substituted phenyl, specifically chlorine or bromine, heterocycle ("het"), what

independently selected from

<formula> formula see original document page 4 </formula>

wherein R2 is straight chain or branched alkyl

<formula> formula see original document page 4 </formula>

where R3 is

X is hydrogen or alkoxy, and pharmaceutically acceptable salts thereof.

To the extent that any of the compounds were previously described in PCT / AU2004 / 000866 as antiviral agents, they are excluded from the present invention.

Preferably, the compounds of the invention include the following:

(3-benzoyl) cinnamoylguanidine,

5-methyl-2-naphthoylguanidine,

3- (indan-4-yl) propenoylguanidine,

5-bromo-6-methoxy-2-naphthoylguanidine,

5-thiophen-3-yl-2-naphthoylguanidine,

5- (1-methylpyrazol-4-yl) 2-naphthoylguanidine,

2,3-methylenedioxycinamolguanidine,

(1-methoxy-2-naphthoyl) guanidine,

(3-methoxy-2-naphthoyl) guanidine,

(5-bromo-2-naphthoyl) guanidine,

(1,4-dimethoxy-2-naphthoyl) guanidine, (6- (3-thienyl) 2-naphthoyl) guanidine, (6-methyl-2-naphthoyl) guanidine, (5-phenyl-2-naphthoyl) guanidine, ( 5- (thien-2-yl) -2-naphthoyl) guanidine, (5- (1,3,5-trimethylpyrazol-4-yl) -2-naphthoyl) guanidine, (5- (1-isobutyl-1 H- pyrazoyl-4-yl) naphthoyl) guanidine, (5- (3-furyl) -2-naphthoyl) guanidine, (5-cyclopropyl-2-naphthoyl) guanidine, (5-chloro-2-naphthoyl) guanidine, (6- Guanidinium (1-methylpyrazol-4-yl) -2-naphthoyl) acetate, (5- (2,6-dimethoxypyridin-3-yl) -2-naphthoyl) guanidine, (5- (2-chlorophenyl) -2- naphthoyl) guanidine, (5- (4- (acetylamino) phenyl) -2-naphthoyl) guanidine, (5- (3- (acetylamino) phenyl) -2-naphthoyl) guanidine, (5- (4 - ((methylsulfonyl) amino) phenyl) -2-naphthoyl) guanidine, and pharmaceutically acceptable salts thereof.

The amine or imine groups of the guanidyl moiety of the compounds of Formula I may be present in any conventional form used for the provision of such compounds. For example, they may be present as the free base, a hydrate, an organic or inorganic salt, or combinations thereof.

Preferably, the compounds of the invention have anti-viral activity, and are capable of reducing, retarding or otherwise inhibiting viral growth and / or replication.

Examples of preferred viruses against which the compounds of the present invention are active are viruses from the Flavivirus or Lenti virus families. More preferably, the virus is Hepatitis C virus (HCV) 1 Human Immunodeficiency Virus (HIV), or dengue virus. More preferably, the ovirus is HCV, HIV-1, HIV-2.

According to a second aspect, the present invention provides a pharmaceutical composition comprising a compound according to a first aspect, and optionally one or more pharmaceutically acceptable derivatives or vehicles. The active compounds may be present in the form of any suitable salt, adduct, in anhydrous or solvated forms.

In one embodiment, the compositions of the invention further comprise one or more known compounds or molecules having antiviral activity. The known antiviral compounds can be selected from the group consisting of Vidarabine, Acyclovir, Ganciclovir, Valganciclovir, Valacyclovir, Cidofovir, Famciclovir, Ribavirin, Amantadine, Rimantadine 1 Interferon, Oseltamivir, Rupivizumab nucleotide inhibitor, Rimantadine anamide, (NRTI) such as Zldovudine, Didanosine1 Zalcitabine1 Stavudine1 Lamivudine and Abacavir1 non-nucleoside reverse transcriptase inhibitors (NNRTI) 1 such as Ne-virapine, Delavirdine and Efavirenz1 protease inhibitors such as Saavina-Navirinavin1 and other known antiviral compounds and preparations.

According to a third aspect, there is provided a method for reducing, retarding, or otherwise inhibiting the growth and / or replication of a virus comprising contacting a cell infected with or exposed to said virus, with a compound according to the first aspect.

According to a fourth aspect, a method is provided for preventing infection of a cell exposed to a virus, comprising contacting said cell with a compound according to the first aspect.

According to a fifth aspect of the invention, there is provided a method for the therapeutic or prophylactic treatment of a subject exposed or infected with a virus, comprising administering to said individual a compound according to the first aspect.

Preferably, the virus is from the Flavivirus and Lenti-virus families. More preferably, the virus is Hepatitis C virus (HCV), Human Immunodeficiency Virus (HIV), or dengue virus. More preferably, the virus is HCV, HIV-1 and HIV-2.

Preferably, the individual supporting therapeutic or prophylactic treatment is a mammal, such as, but not limited to, a human, primate, farm animal (e.g., sheep, cow, horse, donkey, porco), domestic animal (eg dog, cat), laboratory test animal (eg mouse, rabbit, rat, guinea pig, hamster), or captive wild animal (eg fox, deer). individual is a primate. More preferably, the individual is a human.

Preferably, the pharmaceutical composition may additionally comprise one or more known antiviral compounds or molecules. The known antiviral compounds may be selected from the group consisting of Vidarabila, Acyclovir, Ganciclovir, Valganciclovir, Valacy-clovir, Cidofovir, Famciclovir, Ribavirin, Amantadine. , Rimantadine, Interferon, Oseltamivir, Palivizumab, Rimantadine, Zanamivir, nucleoside analog reverse transcript (NRTI) inhibitors such as Zldovudine, Didanosine, Zalcitabine, Stavudina1 Lamivudine and Abacavir, non-transcriptase reverse inhibitors (NNRTI), such as Nevirapine, Delavirdine and Efavirenz, protease inhibitors, such as Saquinavir, Ritonavir, Indinavir, Nelfinavir, Amprenavir, are other known antiviral compounds and preparations.

In the case of any inconsistencies in the present report between the named compounds and the spiruural formula, the structural formula is to be preferred.

Unless the context clearly requires otherwise, throughout the description and claims, the words 'comprising', 'comprising', and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is, in the sense of "including but not limited to".

BRIEF DESCRIPTION OF DRAWINGS

Figure 1. Inhibition of HIV-1Ea-1 BIT compound and cell cytotoxicity in primary human macrophages. "% VC" dose response curves represent the "percent control virus growth rate" figures calculated based on mean virus (triple well) levels in compound containing wells (in decreased concentration) compared to controls (none compound). HIV-1 levels were determined using p24 ELISA, and converted to percent control values based on p24 virus levels detected in control wells containing no compound. The "% viability" curve was calculated from data (DD560nm generated from MTT assays as a measure of cell viability. OD (mean triplicate wells) values were converted to a percentage of controls (not containing compound). ) The 50% level is indicated by the horizontal lineage to allow the estimation of IC50 and TC50 values.

FIGURE 2. CELL CYTOTOXICITY. COMPOUNDS FORMATTED FOR CYTOTOXICITY IN A WIDE CONCENTRATION RANGE

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based, in part, on the surprising observation that certain substituted acylguanides have antiviral activity against a broad range of viruses, including those from the Lentivirus and Flavivirus families.

The present invention relates to the compound of Formula I:

<formula> formula see original document page 8 </formula>

in which

R1 is phenyl, substituted phenyl, naphthyl, substituted naphthyl, or R1 is selected from

<formula> formula see original document page 8 </formula>

or

<formula> formula see original document page 8 </formula> η is 1, 2, 3 or 4;

<formula> formula see original document page 9 </formula>

F is independently halogen, alkyl, halo or polyhaloalkyl;

Q is independently hydrogen, especially methoxy alkoxy, especially methyl alkyl, cycloalkyl, thienyl, furyl, pyrazoyl, substituted pyrazoyl, pyridyl, substituted pyridyl, phenyl, substituted phenyl, chlorine or bromine halo, heterocycle ("het"), what

<formula> formula see original document page 9 </formula>

independently selected from

<formula> formula see original document page 9 </formula>

wherein R2 is straight chain or branched alkyl

<formula> formula see original document page 9 </formula>

X is hydrogen or alkoxy, and pharmaceutically acceptable salts thereof.

To the extent that any of the compounds were previously described in PCT / AU2004 / 000866 as antiviral agents, they are excluded from the present invention.

In particular, useful compounds may be selected from the following:

(3-benzoyl) cinnamoylguanidine comprising the structure

<formula> formula see original document page 10 </formula>

2,3-methylenedioxycinnamoyl guanidine comprising the structure

<formula> formula see original document page 10 </formula>

5-methyl-2-naphthoylguanidine comprising the structure

<formula> formula see original document page 10 </formula>

3- (indan-4-yl) propenoylguanidine comprising the structure

<formula> formula see original document page 10 </formula>

5-bromo-6-methoxy-2-naphthojuanidine comprising the structure

<formula> formula see original document page 10 </formula>

5-thiophen-3-yl-2-naphthoylguanidine comprising the structure

<formula> formula see original document page 10 </formula> 5- (1-methylpyrazol-4-yl) 2-naphthoylguanidine comprising the structure

<formula> formula see original document page 11 </formula>

(1-methoxy-2-naphthoyl) guanidine comprising the structure

<formula> formula see original document page 11 </formula>

(3-methoxy-2-naphthoyl) guanidine comprising the structure

<formula> formula see original document page 11 </formula>

(5-bromo-2-naphthoyl) guanidine comprising the structure

<formula> formula see original document page 11 </formula>

(1,4-dimethoxy-2-naphthoyl) guanidine comprising the structure

<formula> formula see original document page 11 </formula> (6- (3-thienyl) 2-naphthoyl) guanidine comprising the structure

<formula> formula see original document page 12 </formula>

(6-methyl-2-naphthoyl) guanidine comprising the structure

<formula> formula see original document page 12 </formula>

(5-phenyl-2-naphthoyl) guanidine comprising the structure

<formula> formula see original document page 12 </formula>

(5- (thien-2-yl) -2-naphthoyl) guanidine comprising the structure

<formula> formula see original document page 12 </formula>

(5- (1,3,5-trimethylpyrazol-4-yl) -2-naphthoyl) guanidine comprising the <formula> formula see original document page 13 </formula>

(5- (1-Isobutyl-1H-pyrazoyl-4-yl) naphthoyl) guanidine comprising the structure

<formula> formula see original document page 13 </formula>

(5- (3-furyl) -2-naphthoyl) guanidine comprising the structure

<formula> formula see original document page 13 </formula>

(5-cyclopropyl-2-naphthoyl) guanidine comprising the structure

<formula> formula see original document page 13 </formula>

(5-chloro-2-naphthoyl) guanidine comprising the structure <formula> formula see original document page 14 </formula>

Guanidinium (6- (1-methylpyrazol-4-yl) -2-naphthoyl) acetate,

<formula> formula see original document page 14 </formula>

(5- (2,6-dimethoxypyridin-3-yl) -2-naphthoyl) guanidine

<formula> formula see original document page 14 </formula>

(5- (2-chlorophenyl) -2-naphthoyl) guanidine

<formula> formula see original document page 14 </formula>

(5- (4- (acetylamino) phenyl) -2-naphthoyl) guanidine <formula> formula see original document page 15 </formula>

(5- (3- (acetylamino) phenyl) -2-naphthoyl) guanidine

<formula> formula see original document page 15 </formula>

(5- (4 - ((methylsulfonyl) amino) phenyl) -2-naphthoyl) guanidine

<formula> formula see original document page 15 </formula>

and pharmaceutically acceptable salts thereof. The amine or imine groups of the guanidyl portion of the compounds of Formula I may be present in any convenient form used for the provision of such compounds. For example, they may be present as a free base, a hydrate, an organic or inorganic salt, or combinations thereof. The methods developed for classifying the compounds of the present invention for antiviral activity are described in detail in PCT / AU2004 / 000866, incorporated in their entirety. by reference.

Reference to "HIV", "HCV" and "Dengue Virus" and the like should be understood as referring to any strain of HIV, HCV or Dengue virus, and including homologs and mutants.

Reference to the "functional activity" of a virus should be understood as referring to any or more functions that a virus performs or is involved in.

Reference to "viral replication" should be understood to include any one or more stages or aspects of the viral life cycle, such as inhibition of virion assembly or release. Accordingly, the method of the present invention involves the mediation of viral replication via the induction of a cascade of steps leading to the mediation of any or more aspects or stages of the viral life cycle.

Reference to a "cell" infected with a virus should be understood as referring to any cell, prokaryotic or eukaryotic, that has been infected with a virus. This includes, for example, immortal or primary cell lines, bacterial cultures, and cells in situ.

It will be appreciated by those skilled in the art that the compounds of the invention may be administered in the form of a composition or formulation comprising pharmaceutically acceptable carriers, and / or excipients.

The pharmaceutical compositions of the invention may further comprise one or more known antiviral compounds, or molecules. Preferably, the known antiviral compounds are selected from the group consisting of Vidarabine, Acyclovir, Ganciclovir, Valgan-cyclovir, Valacyclovir, Cidofovir, Famciclovir, Ribavirin, Amantadine, Rimanta-dina, Interferon, Oseltamivir, Palivizinase, Rimantavirase, Transmantadine Zmant, nucleoside analogues (NRTI) such as Zldovu-dine, Didanosine, Zalcitabine, Stavudine, Lamivudine and Abacavir, non-nucleoside reverse transcriptase inhibitors (NNRTI) such as Nevirapine, Delavirdine and Efavirenz, protease inhibitors such as Saquinavir1 navir, Indinavir, Nelfinavir1 Amprenavir, and other known antiviral compounds and preparations.

The viral inhibition individual is a mammal, such as, but not limited to, a human, primate, farm animal (e.g., ram, cow, horse, donkey, pig), domestic animal (e.g., dog, cat), laboratory test animal (eg mouse, rabbit, rat, guinea pig, hamster), or captive wild animal (eg fox, deer). Preferably, the individual is a primate. More preferably, the individual is a human.

The method of the present invention is useful in the treatment and prophylaxis of viral infection, such as, for example, HIV, HCV, Dengue, and other viral infections. For example, antiviral activity may be affected in individuals known to be infected with HIV to prevent HIV replication, thereby preventing the attack of AIDS. Alternatively, the method of the present invention may be used to reduce serum viral load, or to alleviate symptoms of viral infection. This concept applies to any viral confection.

The method of the present invention may be particularly useful at earlier stages of viral infection to prevent the establishment of a viral reservoir in affected cells, or as prophylactic treatment to be applied immediately before, or for a period after exposure to a possible source of virus.

Reference here to "therapeutic" and "prophylactic" is to be considered in their broader contexts. The term "therapeutic" does not necessarily imply that a mammal is treated until full recovery. Similarly, "prophylactic" does not necessarily mean that the individual will not eventually encounter a disease condition. Accordingly, therapy and prophylaxis include amelioration of the symptoms of a particular condition or prevention or otherwise reducing the risk of developing a particular condition. The term "prophylaxis" may be considered to reduce the severity of onset of a particular condition. Therapy may also reduce the severity of an existing condition, or the frequency of acute attacks.

In accordance with the methods of the present invention, more than one compound or composition may be co-administered with one or more other compounds, such as known antiviral compounds or molecules.

By "co-administered" is meant simultaneous administration in the same formulation, or in two different formulations, via the same or different routes, or sequential administration by the same or different routes.

By "sequential" administration, a time difference of seconds, minutes, hours or days between the administration of the two or more separate compounds is significant. The subject's antiviral compounds may be administered in any order.

Routes of administration include, but are not limited to, intravenous, intraperitoneal, subcutaneous, intracranial, intradermal, intramuscular, intraocular, intrathecal, intracerebral, intranasal, transmucosal, or orally, rectally, gout, implant and implant infusion. . Intravenous routes are particularly preferred.

Compositions suitable for injectable use include sterile aqueous solutions (where water-soluble), and sterile powders for the temporary preparation of sterile injectable solutions. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g. glycerol, propylene glycol and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. Prevention of the action of microorganisms may be provided by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal and the like. In many cases, it will be preferable to include isotonic agents, for example sugars or sodium chloride. Prolonged absorption of the injectable compositions may be provided by use in the compositions of absorption-delaying agents, for example aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by incorporating active compounds in the required amount in the appropriate solvent with various other ingredients listed above as required, followed, for example, by filter sterilization or other sterilization by appropriate means. Dispersions are also contemplated, and these may be prepared by incorporating the various stabilized active ingredients into a sterile vehicle containing the basic dispersion medium and the other ingredients required from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, a preferred method includes the vacuum drying and freeze-drying technique, which produces a powder of the active ingredient plus any additional ingredient of a previously sterile filtered solution.

When the active ingredients are adequately protected, they may be administered orally, for example, with an inert diluent, or with an assimilable edible carrier, or they may be enclosed in soft or hard shell gelatin capsules, or can be compressed into tablets. For oral therapeutic administration, the compound may be incorporated with excipients, and used in the form of digestible tablets, buccal tablets, tablets, capsules, elixirs, syrups, pastilles and the like. Such compositions and preparations should contain at least 0.01 wt.%, More preferably 0.1 wt.%, Even more preferably 1 wt.% Of active compound. The percentage of the compositions and preparations may, of course, be varied, and may conveniently be from about 1 to about 99%, more preferably from about 2 to about 90%, even more preferably from about 5 to about 80%. The amount of active compound in such a therapeutically useful composition is such that a suitable dosage will be obtained Preferred compositions or preparations according to the present invention are prepared so that an oral dosage unit form contains between about 0.1 g and 2000 mg of active compound.

Tablets, tablets, pills, capsules and the like also contain the components as listed hereinafter. A binder, such as gum, acacia, cornstarch or gelatin; excipients, such as dicalcium phosphates; a disintegrating agent such as cornstarch, potato starch, alginic acid and the like; a lubricant, such as magnesium stearate; and a sweetening agent such as sucrose, lactose and saccharin may be added, or a flavoring agent such as peppermint, wintergreen oil, or cherry flavoring. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier. Various other materials may be present as coatings or otherwise modify the physical form of the dosage unit. For example, tablets, pills or capsules may be coated with varnish, sugar or both. A syrup or elixir may contain the active compound, sucrose as a sweetening agent, methyl and propyl parabens as preservatives, a colorant, and flavoring such as cherry or orange aroma. Any material used in the preparation of any dosage unit form must be pharmaceutically pure, substantially non-toxic in the amounts employed. In addition, the active compound (s) may be incorporated into sustained release preparations and formulations.

The present invention also extends to forms suitable for topical application, such as creams, lotions and gels. In such forms, anticoagulant peptides may need to be modified to allow surface barrier penetration.

Procedures for the preparation of unit dosage forms and topical preparations are readily available to those skilled in the art from texts such as Pharmaceutical Handbook AMartindale Companion Volume Ed. Ainley Wade Nineteenth Edition The Pharmaceutical Press London, CRC Handbook of Chemistry and Physies Ed.Robert C. Weast Ph.D. CRC Press Inc .; Goodman and Gilman's; The Pharmaeological basis of Therapeuties. Ninth Ed. MeGraw Hill; Remington; and The Science and Practice of Pharmacy. Nineteenth Ed. Ed. Alfonso R. Gen. Naro Maek Publishing Co. Easton Pennsylvania.

Pharmaceutically acceptable carriers and / or diluents include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic, and retarding agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except unless any conventional means or agent is incompatible with the active ingredient, use of these therapeutic compositions is contemplated. Additional active ingredients may also be incorporated into the compositions.

It is especially advantageous to formulate parenteral compositions in unit dosage form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically distinct units suitable as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined amount of active material calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for the novel dosage unit forms of the invention are dictated by and directly dependent upon (a) the unique characteristics of the active material and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the composition technique.

Actual amounts contemplated by the present invention will vary depending upon the severity of the condition, and the health and age of the recipient. Generally, effective amounts may range from 0.01 ng / kg body weight to about 100 mg / kg body weight.

Alternative amounts include about 0.1 ng / kg body weight at about 100 mg / kg body weight, or about 1 ng / kg body weight at about 80 mg / kg body weight.

The individual from viral inhibition is generally a mammal, such as, but not limited to, a human, primate, farmed animal (eg, sheep, cow, horse, donkey, pig), domestic animal (e.g. , dog, cat), laboratory test animal (eg, mouse, rabbit, rat, guinea pig, hamster), or captive wild animal (eg, fox, deer). Preferably, the individual is a human or primate. More preferably, the individual is a human.

The present invention will now be described in greater detail with reference to specific, but not limiting, examples describing synthetic protocols, viral inhibition and other antiviral properties of the compounds of the present invention. Synthesis and classification of compounds having antiviral activity may be achieved by the range of methodologies described herein, or described in greater detail in PCT / AU2004 / 000866, incorporated in their entirety herein by reference.

It is to be understood, however, that the description of specific procedures, compounds and methods, is included only for the purpose of exemplifying the present invention. It should not be understood in any way as a restriction on the broad description of the invention as set forth above.

EXAMPLES

The antiviral activity of all compounds of the present invention may be, and has been achieved using the methods described herein, or described in detail in PCT / AU2004 / 000866, incorporated in its entirety herein by reference. Additionally, methods for synthesizing the generic and specific compounds of the invention described herein described in the referenced publications or otherwise known to those skilled in the art may be used to prepare all compounds of the present invention.

More specifically, acylguanidines may be synthesized by a variety of methods including reaction of guanidine (generally generated in situ from "its hydrochloride salt) with a suitably activated derivative of a carboxylic acid. Examples include:

(i) synthesis of acid chlorides, exemplified by Yamamoto etal., Chem. Pharm. Bull., 1997, 45, 1282.

(ii) synthesis of single esters, exemplified by US 2,734,904.

(iii) carboxylic acid synthesis via in situ activation by carbonyl diimidazole, exemplified by US Patent 5,883,133.

The carboxylic acid precursors required for the preparation of the acylguanidines described herein were obtained by a variety of different methods. A large number of the substituted cinnamic acids are commercially available. In addition, numerous procedures for the synthesis of substituted cinnamic acids and their simple esters are well described in the art, including

i) The reaction of malonic acid with an aromatic aldehyde and base (the Doebner Condensation), described in Chemical Reviews, 1944, 35, 156, and references contained therein.

ii) The reaction of acetic anhydride with an aromatic aldehyde and base (the Perkin Reaction), described in Organic Reaction, 1942,1, 210, and references contained therein.

iii) The reaction of acrylic acid and simple esters thereof with an aromatic halide or aromatic triflate using palladium catalyst (theHeck Reaction), described in Organic Reactions, 1982, 28, 345, and references contained therein.

iv) The reaction of a trialkyl phosphonoacetate with an aromatic aldehyde and base (the Horner-Emmous Reaction), described in Organic Reaeti-ons, 1977, 25, 73, and references contained therein.

A number of single halo, hydroxy, and alkoxysubstituted naphthoic acids are either commercially available or known in the art, and these provide the starting materials for substituted naphthoylguanidines.

Naphthoic acids that are substituted with alkyl, cycloalkyl, aryl, and heterocyclic groups can often be prepared by reacting a halonaftoic acid with an appropriate organometallic reagent using a transition metal catalyst. One such variant of this methodology that was used to prepare a number of the substituted naphthoic acids used as precursors to the naphthoylguanidines described herein was the palladium-catalyzed carbon-carbon bond formation reaction between bromonaphoic acids and a suitable substituted bromic acid (or ester). boronate) which is widely known in the art as Suzuki coupling (described in Chemical Review, 1995, 95, 2457, references in this). The reaction has broad applicability, and can be used in a range of substituted halonaphthalenes which can then be designed to introduce or not mask the required carboxylic acid group.

1. General Synthetic Methodology

1.1 General Procedure A - Preparation of Aryl Triflates

To a solution of phenol (10 mmol) in pyridine (7 mL) at 0 ° C, trifluoromethanesulfonic anhydride (11 mmol, 1eq.) Was slowly added. The resulting mixture was stirred at 0 ° C for an additional 5 minutes before being allowed to warm to room temperature, and stirred until TLC analysis showed that the starting phenol had been consumed. The mixture was then poured into water and extracted with ethyl acetate (x3). The combined extracts were washed sequentially with water, 1M aqueous hydrochloric acid, water and brine, then dried (MgSO4) 1 and concentrated in vacuo to give crude product. The crude products were chromatographed on silica gel. Elution with a mixture of ethyl acetate / hexane gave desired aryl triflates, generally as colorless oils.

1.2 General Procedure B - Cinnamate Esters Via Triflate Check Reaction

A mixture of phenyl triflate (10 mmol), methyl acrylate (14 mmol, 1.4 eq.), Triethylamine (40 mmol, 4 eq.), And dichlorobis (triphenylphosphine) palladium (0.3 mmol, 0.03 eq) in Dimethylformamide (30 mL) was heated to 90 ° C. The reaction was monitored by GC / MS and fresh batches of methyl acrylate (1 eq), triethylamine (2 eq) and palladium catalyst (0.03 eq) were added as required in an effort to force the reaction to completion. The mixture was then poured into water and extracted with a 1: 1 mixture of diethyl ether / hexane (x3) .The combined extracts were washed with water, then brine, dried (MgSO4), filtered through a desiccant pad. The filtrate was concentrated in vacuo to give the crude product as an oil The crude products were chromatographed on silica gel Elution with a mixture of ethyl acetate / hexane gave the desired methyl cinnamates, generally as colorless oils.

1.3 General Procedure C - Cinnamate Esters Via Bromide Dekeck Reaction

Aryl bromide (10 mmols), palladium acetate (0.1 mmoles, 0.01eq) and tri-o-tolylphosphine (0.4 mmols, 0.04 eq) were added to the dereation flask, and purged with nitrogen. To this, ethyl macetate (12.5 mmol, 1.25 eq), triethylamine (12.5 mmol, 1.25 eq), and dimethylformamide (1 mL) were then added, and the mixture was heated to 100 ° C. The reaction was monitored by GC / MS and fresh batches of palladium acetate (0.01eq) tri-o-tolylphosphine (0.04eq), methyl acrylate (1.25eq) and triethylamine (1.25eq) were added as follows. required in an effort to force the reaction to completion. The mixture was poured into water and extracted with a 1: 1 mixture of diethyl ether / hexane (x4). The combined extracts were washed with water, then brine, dried (MgSO 4), filtered through a silica gel pad, and the filtrate was concentrated in vacuo to afford the crude product. The crude products were chromatographed on silica gel. Elution with a mixture of ethyl acetate / hexane gave the desired methyl cinnamates, generally as colorless oils.

1.4 General Procedure D - Cinnamate Esters Via Horner-Emmons Reaction

A solution of triethyl phosphonoacetate (13 mmol, 1.3 eq) in anhydrous tetrahydrofuran (10 mL) was added over 5 minutes to a suspension of sodium hydroxide (14.3 mmol, 1.4 eq) in anhydrous tetrahydrofuran (10 mL) at 0 ° C under nitrogen. The mixture was then stirred at 0 ° C for 20 minutes. A solution of benzaldehyde (10 mmol) in teirahldrofuran (15 µmL) was then added over 10 minutes at 0 ° C. The mixture was stirred at 0 ° C for an additional 30 minutes before being allowed to stir at room temperature until GC / MS or TLC analysis showed that benzaldehyde starting material had been consumed. Typically, reactions were allowed to stir at room temperature overnight to ensure complete consumption of the starting aldehyde. The mixture was poured into water, the organic layer was separated, and the aqueous layer was extracted with ethyl (x3) comacetate. The combined organic extracts were washed with water, then brine, dried (MgSO4) 1 and concentrated in vacuo to give crude product. The crude products were chromatographed on silyl gel. Elution with a mixture of ethyl acetate / hexane gave the desired methylcinnamates, generally as colorless oils.

1.5 General Procedure E - Preparation of 5-Phenylpenta-2,4-Dienoic Esters

A solution of anhydrous triethyl 4-phosphonocrotonate (26 mmol, 1.4 eq) in tetrahydrofuran (10 mL) was added over 5 minutes to a suspension of sodium hydroxide (28 mmol, 1.4 eq, 60% suspension in oil). ) in anhydrous tetrahydrofuran (15 mL) at 0 ° C under nitrogen. The mixture was then stirred at 0 ° C for 20 minutes. A solution of benzaldehyde (20 mmol) in tetrahydrofuran (10 mL) was then added over 10 minutes at 0 ° C.

The mixture was stirred at 0 ° C for an additional 30 minutes, and then allowed to stir at room temperature until GC / MS or TLC analysis showed that the starting aldehyde had been consumed. The reaction mixture was poured into water, the organic layer was separated, and the aqueous layer was extracted with ethyl acetate (x3). The combined organic extracts were then washed with water, then brine, dried (Mg-SO4), and concentrated in vacuo to give the crude ethyl ester as an oil. The crude products were chromatographed on silica gel. Elution with a mixture of ethyl acetate / hexane gave the desired ethyl esters as colorless oils.

1J3 General Procedure F - Ester Hydrolysis

A solution of the ester (10 mmol) in melanol (50 mL) and water (5 mL) was treated with a 6M aqueous potassium hydroxide solution (20 mmol), and the mixture was heated under reflux until TLC analysis showed. no starting material was present anymore (usually 2-3 hours). The mixture was then poured into water (50-200 mL), and acidified with concentrated hydrochloric acid to approximately pH 2. The resulting carboxylic acid was collected by filtration, washed with water, and dried over all night under high vacuum.

1.7 General Procedure G - Suzuki Reactions of Bro-Monaptoic Acids

Bromo-2-naphthoic acid (2 mmols), appropriate boronic acid (or boronate ester), tetrakis (triphenylphosphine) palladium (0) (0.1 mmols), and solid sodium carbonate (6.8 mmols) were added to the reaction flask which was then purged with nitrogen. Acetonitrile (6 ml) and water (2.5 ml) were added, and the mixture was heated under reflux with vigorous stirring until the starting bromo-2-naphthoic acid was consumed. The reaction mixture was then partitioned between toluene (50 mL) and 0.5M sodium hydroxide solution (100 mL). The aqueous layer was washed with toluene (to remove any triphenylphosphine, 3 x 20 mL), then acidified to pH 1 with concentrated hydrochloric acid. The naphthoic acid derivatives were extracted into ethyl acetate (4 χ 20 mL). The combined ethyl acetate extracts were washed with water (3 x 20 mL) and brine, then dried (MgSO4), filtered and concentrated. The residue was analyzed by 1 H NMR, and chromatographed on silica gel (if required).

1.8 General Procedure H - Preparation of Acylguanidines

To a suspension / solution of carboxylic acid (10 mmols) in dichloromethane (30 mL) containing a drop of dimethylformamide was added oxalyl chloride (12 mmols, 1.2 eq) which made the solution become effervescent. . After stirring for 2 hours, the resulting solution was evaporated to dryness under reduced pressure. The residue was dissolved in dry tetrahydrofuran (30 mL) and added to a solution of guaminid hydrochloride (50 mmols, 5.0 eq) in 2M aqueous sodium hydroxide (30 mL). The reaction was stirred at room temperature for 1 hour and then the tetrahydrofuran layer was separated. The aqueous layer was extracted with chloroform (100 mL), followed by ethyl acetate (100 mL), and the combined organic layers evaporated under reduced pressure. The resulting residue was partitioned between chloroform (200 mL) and 2M aqueous sodium hydroxide (100 mL), and the organic layer was separated and dried (Na 2 SO 4). The solution was filtered and evaporated under reduced pressure to the point where a solid began to precipitate. At this point, hexanes were added, causing precipitation of the product which was collected by filtration and dried under high vacuum.

2. EXPERIMENTAL EXAMPLES OF SYNTHESIS EXAMPLE 1: 4-HYDROXYINDAN

4-Aminoindan (3.0 g) was added to a solution of concentrated sulfuric acid (2.4 mL) in water (15 mL). More water (15 mL) was added, and the mixture cooled to 5 ° C. A solution of sodium nitrite (1.71 g) in water (4.5 mL) was added portion by portion to the mixture while weekly at a temperature below 5 ° C. After addition was complete, the mixture was allowed to warm to room temperature, and urea (0.29 g) was added. The mixture was stirred for another 5 minutes before being heated at 45 ° C for 30 minutes. The mixture was then cooled to room temperature and extracted with ethyl acetate. The combined organic extracts were washed with 2M aqueous sodium hydroxide (2 χ 100 mL), and these aqueous extracts were then acidified with hydrochloric acid and extracted with ethyl acetate (3 χ 100 mL). The combined organic extracts were then washed with brine and dried (Na 2 SO 4) before being concentrated in vacuo. The resulting crude product was chromatographed on silica gel. Elution with ethyl acetate / hexane (1: 7) gave 4-hydroxyindan as an orange oil (1.0 g).

EXAMPLE 2: 4-INDANY TRIFLATO

To a solution of 4-hydroxyindan (1.2 g, 8.9 mmol) in pyridine (5 mL) at 0 ° C was slowly added trifluoromethanesulfonic anhydride (1.6 mL, 9.8 mmol). The resulting mixture was stirred at 0 ° C for 5 minutes before being allowed to warm up to room temperature, and then stirred for 45 minutes. The mixture was then poured into water and extracted with ethyl acetate (3 x 25 mL). The combined extracts were subsequently washed with water, 1M aqueous hydrochloric acid, water and brine, then dried (Na 2 SO 4), and concentrated in vacuo to give triflatobruto as an orange oil (2.13 g, 89%).

EXAMPLE 3: 3- (INPAN-4-IL) ACRYLATE METHYL

A mixture of crude 4-indanyl triflate (2.13 g, 8.0 mmol), methylacrylate (1.101 mL, 11.2 mmol), triethylamine (4.4 mL, 32 mmol, 4 eq) and dichlorobis (triphenylphosphine) Palladium (170 mg, 0.24 mmol) in dimethylformamide (15 mL) was heated at 85 ° C for 71 hours. A small aliquot was removed and operated for GC / MS analysis which revealed that a significant amount of starting material was still present. Additional methyl acrylate (0.7 mL), triethylamine (2 mL), and palladium catalyst (170 mg) were added, and the mixture was heated for another 24 hours. The mixture was then poured into water, extracted with ethyl acetate, and the organic extracts were washed with water, then brine, dried (Na 2 SO 4), and concentrated in vacuo to give crude product. as an oil (2.4 g). The product was chromatographed on silica gel. Elution with ethyl acetate / hexane (1:19) gave the starting triflate (812 mg, 38%) as a colorless oil, followed by the desired methyl-3- (indan-4-yl) acrylate as a brown oil (880 mg, 54%).

EXAMPLE 4: 3-BROMOBENZOPHENONE METHYL

To a mixture of 3-bromobenzophenone (5.0 g, 19 mmol), palladium acetate (215 mg, 0.958 mmol) and tri-o-tolylphosphine (290 mg, 0.953 mmol) was added triethylamine (3.3 mg). mL, 45 mmol), toluene (4 mL), and methylacrylate (2.2 mL, 27 mmol). The mixture was heated at 100 ° C for 18 hours at which time TLC analysis showed that the reactions were still incomplete. Additional portions of palladium acetate (215 mg, 0.958 mmol), tri-o-tolylphosphine (290 mg, 0.953 mmol), triethylamine (3.3 mL, 45 mmol), and methylacrylate (2.2 mL, 27 mmol) were added. added, and the mixture was heated to 110 ° C for another 18 hours. After cooling to room temperature, the mixture was poured into water, and extracted with ethyl acetate (3 χ 100 ™ rnL). The combined organic extracts were washed sequentially with water and brine, then dried (Na 2 SO 4), and concentrated to a brown oil (5.3 g). The oil was chromatographed on silica gel. Elution ethyl acetate / hexane (1: 9) afforded methyl-3-benzoylcinnamate (4.6 g, 91%) as a yellow solid.

EXAMPLE 5: 3-BENZOYLAMINIC ACID

Aqueous 5M potassium hydroxide (10 mL, 50 mmol) was added to a solution of methyl-3-benzoylcinnamate (2.5 g, 9.4 mmol) in methanol (20 mL), and the mixture was stirred. at room temperature for 18 hours.

The mixture was concentrated and acidified to pH 1 using 1M hydrochloric acid. The resulting precipitate was collected by filtration and dried under vacuum to give 3-benzoyl cinnamic acid (2.2 g, 93%) as a yellow solid.

EXAMPLE 6: 5- (1-METHYL-1H-PIRAZOL-4-lü-2-NAPHTHIC ACID

A mixture of 5-bromo-2-naphthoic acid (2.12 g, 8.44 mmol), 1-methyl-4- (4,4,5,5-tetramethyl-1,3,2-dioxoborolan-2-one il) -1 H-pyrazole (1.84 g, 8.86 mmols), and tetrakis (triphenylphosphine) palladium (0) (502 mg, 0.435 mmols) in a 250 mL round bottom flask was evacuated and purged with nitrogen (in three cycles). Acetonitrile (40 mL) and 2M aqueous sodium carbonate (10mL) were added to the mixture via syringe and the mixture was heated under nitrogen overflow for 22 hours. The reaction mixture was allowed to cool before the addition of 1M aqueous hydrochloric acid (30 mL) and was then extracted with ethyl acetate (3 x 50 mL). The combined organic layers were dried (Na 2 SO 4), filtered, and concentrated in vacuo to afford a crude product (2.98 g after air drying). This crude material was dissolved in hot ethanol (150 mL), and filtered while hot to give a yellow impurity (120 mg). The filtrate was concentrated in vacuo, and the residue was recrystallized from dichloromethane (30 mL) to afford 5- (1-methyl-1H-pyrazol-4-yl) -2-naphthoic acid as a white solid ( 724mg, 34%). A second portion of 5- (1-methyl-1H-pyrazol-4-yl) -2-naphthoic acid (527 mg, 25%) was obtained from the concentrated mother liquors by recrystallization from dichloromethane ( 20 mL).

EXAMPLE 7: 5- (1-METHYL-1 H-FtRAZOL-4-IL-2-NAPHTHYLGUANIDINE

Oxalyl chloride (1.1 mL, 13 mmol) was added to a solution of 5- (1-methyl-1H-pyrazol-4-yl) -naphthoic acid (1.19 g, 4.71 mmol) in dichloromethane anhydrous (200 mL (which was added portionwise during the reaction to dissolve)) containing dimethylformamide (2 drops) under nitrogen, and the mixture was stirred at room temperature for 4.25 hours. The reaction mixture was then heated for 1 hour at 40 ° C before being concentrated under reduced pressure. The resulting acid chloride was suspended in anhydrous tetrahydrofuran (50 mL), and this mixture was added dropwise to a solution of guanidine hydrochloride (2.09 g, 21.9 mmols) in 2M aqueous sodium hydroxide ( 15 mL, 30 mmol), and the reaction mixture was then stirred for 30 minutes. The organic phase was separated and the aqueous phase extracted with chloroform (3 x 30 mL), followed by ethyl acetate (3 x 30 mL). The combined organic extracts were washed sequentially with 1M aqueous sodium hydroxide (60 mL) and water (40 mL), then dried (Na 2 SO 4), and concentrated in vacuo to give a glassy solid (1.45 g after drying under high vacuum). This solid was dissolved in dichloromethane which was then slowly evaporated to give 5- (1-methyl-1H-pyrazol-4-yl) -2-naphthoylguanidine as a yellow solid (1.15 g, 83%).

EXAMPLE 8: Ethyl 2,3-Methylenedioxycinaminate

Triethylphosphonoacetate (4.05 mL, 20.2 mmol) was added dropwise to a stirred suspension of sodium hydroxide (0.8 g, 20 mmol) in anhydrous tetrahydrofuran (20 mL) at 0 ° C under nitrogen. The mixture was stirred at 0 ° C for 20 minutes. A solution of 2,3-methylenedioxybenzaldehyde (2.50 g, 16.7 mmol) in tetrahydrofuran (10 mL) was added dropwise at 0 ° C. The mixture was stirred for 2 hours, during which time it was allowed to warm to room temperature. The mixture was poured into water (250 mL), and ethyl comacetate (3 x 250 mL) was extracted. The combined organic extracts were then washed with brine, dried (MgSO 4), and concentrated in vacuo. The crude product was chromatographed on silica gel. Elution with ethyl acetate / hexane (1:10) gave ethyl 2,3-methylenedioxycinnamate as a colorless solid (3.50 g, 92%).

EXAMPLE 3. 2.3-Methylenedioxycinamic Acid

A solution of ethyl 2,3-methylenedioxycinnamate (3.40 g) in methanol (25 mL) and water (5 mL) was treated with a solution of depotassium hydroxide (4.3 g) in water (25 mL). The mixture was stirred overnight at room temperature before being concentrated in vacuo to half of its original volume. The concentrate was then acidified with concentrated HCl to give 2,3-methylenedioxycinnamic acid as a colorless solid (2.81 g, 95%), which was collected by filtration, and dried overnight under vacuum.

Example 10: 2,3-Methylenedioxycinoylguanidine

Oxalyl chloride (0.68 mL, 7.8 mmol) was added to a suspension of 2,3-methylenedioxycinnamic acid (500 mg, 2.6 mmol) in dichloromethane (5 mL) containing a drop of dimethylformamide. The mixture was stirred for 2.5 hours, and the resulting solution was evaporated to dryness under reduced pressure. The residue was dissolved in dry tetrahydrofuran (5mL) and added to a solution of guanidine hydrochloride (1.24g, 13mmols) in 2M aqueous sodium hydroxide (8ml). The reaction was stirred at room temperature for 1 hour and chloroform was then added. The resulting precipitate of crude product (100 mg) was collected by filtration. The filtrate was extracted with chloroform (3 x 30 mL) and ethyl acetate ( The combined organic extracts were washed with 2M aqueous sodium hydroxide (20 mL), water (20 mL), dried (NaaSO 4), and concentrated under reduced pressure to give an additional amount of crude product (400mg). The two portions of crude product were combined, suspended in chloroform (10 mL), and stirred vigorously for 20 minutes. The resulting 2,3-methylenedioxycinnamoylguanidine (420 mg) was collected by filtration, and dried under vacuum.

EXAMPLE 11: VIRAL INHIBITION TESTS

As indicated above, the methods used to classify the antiviral activity of the compounds of the present invention have been described in detail in PCT / AU2004 / 000866, incorporated in their entirety herein by reference.

Specific inhibition of HIV-1 growth by the compounds of the invention was tested on primary macrophages in vitro using a method similar to that used to access antibody neutralization (Van-Cott TC et al (1999)). In the present case, laboratory-adapted HIV-1 Ba-L strain was used, which is known to infect macrophages.

11.1 Preparation of macrophages

Peripheral Blood Mononuclear Cells (PBMC) were prepared from healthy donors using portions coated from Australian Red Cross Blood Service (ARCBS). Blood was received the day after donation, and had been depleted of serum and platelets, leaving a small volume (less than 100 mL) of concentrated leukocytes.

Leukocytes were separated from blood cells and granulocytes by density gradient centrifugation (Ficoll Paque), and washed extensively in PBS (without Ca / Mg) to remove platelets. Recovered cells were allowed to adhere to tissue culture flasks (Becton Dickinson) for 2 hours, 37 ° C, 5% CO2 in medium (DMEM (high) 10% AB serum / 50 µg / ml gentamicin / 2mM L-glutamine) over time the myeloid cells formed an adherent monolayer that left unfixed lymphocytes.

Non-adherent lymphocytes were removed by washing with warm PSB + (with Ca / Mg), and were generally discarded. In some cases those cells were recovered and frozen in the medium / 10% DM-SO for use in viral stock infections. Adherent monocytes were recovered by gentle scraping with a plastic cell scraper. Myeloid cells were then placed in 96-well plates at a cell concentration of 1.5 χ 106 / mL, and allowed to fully differentiate over a 14-day period.

11.2 TEST PROTOCOL

Macrophages were allowed to differentiate over a 14-day period, with medium changes as required. On the tenth day, the cells were infected in the presence of decreased concentration of BIT compounds over an O - 10μΜ range. Samples of 25 μΙ_ of culture supernatants were removed from each well on the seventh day, and fresh medium plus compound dilution was added.

11.3 ANALYSIS OF VIRAL INHIBITION

After 7 days, samples with and without compound were analyzed to access the level of virus present in the wells using a p24 ELISA (lnnotest). Virus levels in the samples were converted to "control viral growth percentage" values based on the controls (not containing compound). From this dose response curve (example Figure 1), the IC 50 was calculated and used as a measure of compound anti-HIV activity.

11.4 CYTOTOXICITY

The toxicity of the test compounds to cells in the viral inhibition assay was determined using the MTT assay (Pauwels R, et al. (1988); D1 Brutoz OJ et al. (1999); Joo H. (2003)). This method uses ThiazolylAzul (MTT), which is added to the cell culture (100 µg / well), and incubated for 4-5 hours, while metabolically active living cells convert the chemistry into their intracellular crystalline purple metabolites. . The purple color is measured colorimetrically (570-590 nm) once the cells have been permeabilized using isopropanol acidified containing 10% triton X-100. The most intense color development occurs in the cavities where the metabolizing cells are most numerous.

The measured OD values were converted to "percentage viability" figures based on the controls (not containing compounds), and the value at which 50% cell viability (TC50) was then measured can be estimated. These values were estimated manually from Percent Viability vs. Dose Response Curve Concentration (Figure 1).

11.5 RESULTS

IC50 values for each compound were calculated from the individual experiments performed and shown in figures 1 and 2.

Compound toxicity to cultured cells was accounted for in inhibition experiments, and in separate experiments involving increased compound concentrations.

The Antiviral Index (Al) value was calculated using the formula: AI = [TC50] / [IC50]

Table 1. IC50 and Al values for test compounds against

<table> table see original document page 34 </column> </row> <table> <table> table see original document page 35 </column> </row> <table>

All IC50 values were estimated from the dose response curves shown in Figure 1. TC50 values were estimated in separate experiments (not shown), where compound concentrations were higher.

Additional compounds were tested using the same assay system and dose response curves as described above, and their IC50 was estimated. Table 2 shows a summary of the data obtained.

Table 2: Estimated IC 50 values for additional compounds of the invention

<table> table see original document page 35 </column> </row> <table>

EXAMPLE 12: ANTIVIRAL ACTIVITY OF COMPOUNDS USING THE BACTERIAL BIOSAY METHOD

The bacterial bioassay method used in the present example to test the antiviral activity of the compounds against different viral targets has been described in detail in PCT / 2004/000866, incorporated in its entirety herein by reference. The results of the bacterial bioassay tests are summarized in Table 3 below. Vpu, p7 and M referred to in the table are small membrane proteins encoded by HIV1 HCV and Dengue virus, respectively, which have functional activities that support viral growth and replication.

While the invention has been described with reference to specific embodiments, it will be understood that variations and modifications which they maintain within the principles and spirit of the described invention are also involved.

<table> table see original document page 36 </column> </row> <table> <table> table see original document page 37 </column> </row> <table>

REFERENCES

VanCott TC1 Mascfola JR1 Loomis-Price LD1 Sinangil F, Zito-mersky N1 McNeiI J1 Robb ML, Birx DL1 Barnett S. (1999) J. ViroL73 (6): 4640-50Pauwels R1 Balzarini J, Sneck R, Schols D , HerdewijnP, Desmyter J and De Clerq E. (1988) J. Virolog. Methods. 20: 309-321

D'Brutoz OJ, Shih M-J1 Yiv SH1 Chen C-L, Uckurn FM. (1999) Mol. Hum. Reprod. 5 (5): 421-432

Joo, Hong-Gu, (2003) J. Vet. Sci4 (3): 229-234

Claims (19)

1.- Compound of Formula I: <formula> formula see original document page 39 </formula> where R1 is phenyl, substituted phenyl, naphthyl, substituted naphthyl, or R1 is selected from <formula> formula see original document page 39 </ formula > η is 1, 2, 3 or 4; <formula> formula see original document page 39 </formula> F is independently ^ s5r halogen, alkyl, halo or polyhaloalkyl; Q is independently hydrogen, especially methoxy alkoxy, especially methyl alkyl , cycloalkyl, thienyl, furyl, pyrazoyl, substituted pyrazoyl, pyridyl, substituted pyridyl, phenyl, substituted phenyl, chlorine or bromine halo, heterocycle ("het"), or Q is independently selected from <formula> formula see original document page 39 </formula> <formula> formula see original document page 45 / formula> where R2 is straight or branched alkyl, <formula> formula see original document page 45 / formula> where R3 is <formula> formula see original document page 45 / formula> or eX is hydr hydrogen or alkoxy, and pharmaceutically acceptable salts thereof. A compound according to claim 1 wherein the compound is selected from the group consisting of: (3-benzoyl) cinnamoylguanidine, -5-methyl-2-naphthoylguanidine, -3 (indan-4-yl). ) -propenoylguanidine, -5-bromo-6-methoxy-2-naphthoylguanidine, -5-thiophen-3-yl-2-naphthoylguanidine, -5'C'-trimethylpyrazol-4-yl) 2-naphthoylguanidine, -2.3 -methylenedioxycinamolguanidine, (1-methoxy-2-naphthoyl) guanidine, (3-methoxy-2-naphthoyl) guanidine, (5-bromo-2-naphthoyl) guanidine, (1,4-dimethoxy-2-naphthoyl) guanidine, ( 6- (3-thienyl) 2-naphthoyl) guanidine, (6-methyl-2-naphthoyl) guanidine, (5-phenyl-2-naphthoyl) guanidine, (5- (thien-2-yl) -2-naphthoyl) guanidine, (5- (1,3,5-trimethylpyrazol-4-yl) -2-naphthoyl) guanidine, (5- (1-isobutyl-1H-pyrazoyl-4-yl) naphthoyl) guanidine, (5- ( 3-furyl) -2-naphthoyl) guanidine, (5-cyclopropyl-2-naphthoyl) guanidine, (5-chloro-2-naphthoyl) guanidine, (6- (1-methylpyrazol-4-yl) -2-naphthoyl) guanidinium acetate, (5- (2,6-dimethoxypyridin-3-yl) -2-naphthoyl) guanidine, (5- (2-chlorophenyl) -2-naphthoyl) guanidine, (5- (4- (acetylamino) phenyl) ) -2-naphthoyl) guani dine, (5- (3- (acetylamino) phenyl) -2-naphthoyl) guanidine, (5- (4 - ((methylsulfonyl) amino) phenyl) -2-naphthoyl) guanidine, are pharmaceutically acceptable salts thereof. A compound according to claim 1 or 2, wherein the amine or imine groups of the guanidyl moiety of the compounds of Formula I may be present as the free base, a hydrate, an organic or inorganic salt, or combinations thereof. . A compound according to claim 1 or claim 2, wherein the compound has antiviral activity. A compound according to any one of claims 1 to 3, wherein the compound is capable of reducing, retarding or otherwise inhibiting viral growth and / or replication. A compound according to any one of claims 1a - 4, wherein the antiviral activity is directed against viruses of the Flavirus or Lentivirus families. A compound according to claim 5, wherein the virus is selected from the group consisting of Hepatitis C virus (HCV), Human Immunodeficiency Virus (HIV), and dengue virus. A compound according to claim 6, wherein the virus is selected from the group consisting of HCV, HIV-1, HIV-2, andDengue virus. A compound according to any one of claims 1 to 7, wherein the compound is in the form of a salt, an adduct, or in anhydrous or solvated form. A pharmaceutical composition comprising a compound as defined in any one of claims 1 to 8, optionally in combination with or more pharmaceutically acceptable or adjuvant carriers. The pharmaceutical composition of claim 9, further comprising one or more known antiviral agents. A method for reducing, retarding, or otherwise inhibiting the growth and / or replication of a virus comprising contacting a cell infected with or exposed to said virus with a compound as defined in any one of claims 1. to 8, or with a pharmaceutical composition as defined in claim 9 or 10. A method for preventing infection of a virus-exposed cell, comprising contacting said cell with a compound as defined in any one of claims 1 to 8, or with a pharmaceutical composition as defined in claim 9 or 10. A method for the therapeutic or prophylactic treatment of an individual exposed to or infected with a virus, comprising administering to said individual a compound as defined in any one of claims 1 to 8, or a pharmaceutical composition as defined in claim 9 or 10. . A method for the therapeutic or prophylactic treatment of an individual exposed to or infected with a virus, comprising administering to said individual a compound as defined in any one of claims 1 to 8, or a pharmaceutical composition as defined in claim 9, in together another one or more known antiviral agents. A method according to any one of claims 11 to 14, wherein the virus is selected from the Flavivirus or Lentivirus families. The method of claim 15, wherein the virus is selected from the group consisting of Hepatitis C virus (HCV), Human Immunodeficiency Virus (HIV), and dengue virus. The method according to claim 16, wherein the virus is selected from the group consisting of HCV1 HIV-1, HIV-2, andDengue virus. A method according to any one of claims 11 to 17, wherein the subject supporting therapeutic or prophylactic treatment is a mammal selected from the group consisting of human, primate, farmed animal, and domestic animal, laboratory test animal or captive wild animal.